Literature DB >> 29806576

Whole genome amplification and exome sequencing of archived schistosome miracidia.

Winka Le Clec'h1, Frédéric D Chevalier1, Marina McDew-White1, Fiona Allan2, Bonnie L Webster2, Anouk N Gouvras2, Safari Kinunghi3, Louis-Albert Tchuem Tchuenté4, Amadou Garba5, Khalfan A Mohammed6, Shaali M Ame7, Joanne P Webster8, David Rollinson2, Aidan M Emery2, Timothy J C Anderson1.   

Abstract

Adult schistosomes live in the blood vessels and cannot easily be sampled from humans, so archived miracidia larvae hatched from eggs expelled in feces or urine are commonly used for population genetic studies. Large collections of archived miracidia on FTA cards are now available through the Schistosomiasis Collection at the Natural History Museum (SCAN). Here we describe protocols for whole genome amplification of Schistosoma mansoni and Schistosome haematobium miracidia from these cards, as well as real time PCR quantification of amplified schistosome DNA. We used microgram quantities of DNA obtained for exome capture and sequencing of single miracidia, generating dense polymorphism data across the exome. These methods will facilitate the transition from population genetics, using limited numbers of markers to population genomics using genome-wide marker information, maximising the value of collections such as SCAN.

Entities:  

Keywords:  Exome sequencing; FTA cards; Schistosoma; miracidia; quantitative PCR; whole genome amplification

Mesh:

Substances:

Year:  2018        PMID: 29806576      PMCID: PMC6193844          DOI: 10.1017/S0031182018000811

Source DB:  PubMed          Journal:  Parasitology        ISSN: 0031-1820            Impact factor:   3.234


  40 in total

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Journal:  PLoS Pathog       Date:  2019-10-25       Impact factor: 6.823

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